TGM3 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Transglutaminase 3, Epidermal (TGM3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Transglutaminase 3, Epidermal (TGM3) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

AI893889; E polypeptide; protein-glutamine gamma-glutamyltransferase E; protein-glutamine-gamma-glutamyltransferase; TG E; TG (E) ; TGase E; TGase-3; TGE; transglutaminase E; transglutaminase-3; we.

Catalyzes the calcium-dependent formation of isopeptide cross-links between glutamine and lysine residues in various proteins, as well as the conjugation of polyamines to proteins. Involved in the formation of the cornified envelope (CE), a specialized component consisting of covalent cross-links of proteins beneath the plasma membrane of terminally differentiated keratinocytes. Catalyzes small proline-rich proteins (SPRR1 and SPRR2) and LOR cross-linking to form small interchain oligomers, which are further cross-linked by TGM1 onto the growing CE scaffold (By similarity) . In hair follicles, involved in cross-linking structural proteins to hardening the inner root sheath.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Transglutaminase 3, Epidermal (TGM3) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Transglutaminase 3, Epidermal (TGM3) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Transglutaminase 3, Epidermal (TGM3), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Transglutaminase 3, Epidermal (TGM3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 78-93 83 EDTA plasma (n=5) 83-98 88 heparin plasma (n=5) 97-105 101 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 78-93 83 serum (n=5) 78-93 83 EDTA plasma (n=5) 83-98 88 EDTA plasma (n=5) 83-98 88 heparin plasma (n=5) 97-105 101 heparin plasma (n=5) 97-105 101

Component Amount Anti-Tgm3 Microplate 96 Wells (12 x 8 Well Strips) Tgm3 Lyophilized Standard 2 x 20 ng 100X Biotinylated Tgm3 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-Tgm3 Microplate 96 Wells (12 x 8 Well Strips) Anti-Tgm3 Microplate 96 Wells (12 x 8 Well Strips) Tgm3 Lyophilized Standard 2 x 20 ng Tgm3 Lyophilized Standard 2 x 20 ng 100X Biotinylated Tgm3 Detector Antibody 120 uL 100X Biotinylated Tgm3 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Transglutaminase 3, Epidermal (TGM3) . No significant cross-reactivity or interference between Transglutaminase 3, Epidermal (TGM3) and analogues was observed.